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Sci Rep. 2017 Jan 4;7:39385. doi: 10.1038/srep39385.

Sequence specific sorting of DNA molecules with FACS using 3dPCR.

Sukovich DJ1,2, Lance ST1,2,3, Abate AR1,2,3.

Author information

1
Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, USA.
2
California Institute for Quantitative Biosciences, University of California San Francisco, San Francisco, California, USA.
3
UC Berkeley-UCSF Graduate Program in Bioengineering, University of California San Francisco, San Francisco, CA, USA.

Abstract

Genetic heterogeneity is an important feature of many biological systems, but introduces technical challenges to their characterization. Even with the best modern instruments, only a small fraction of DNA molecules present in a sample can be read, and they are recovered in the form of short, hundred-base reads. In this paper, we introduce 3dPCR, a method to sort DNA molecules with sequence specificity. 3dPCR allows heterogeneous populations of DNA to be sorted to recover long targets for deep sequencing. It is valuable whenever a target sequence is rare in a mixed population, such as for characterizing mutations in heterogeneous cancer cell populations or identifying cells containing a specific genetic sequence or infected with a target virus.

PMID:
28051104
PMCID:
PMC5209659
DOI:
10.1038/srep39385
[Indexed for MEDLINE]
Free PMC Article

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